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Quantification of HIV-based lentiviral vectors: influence of several cell type parameters on vector infectivity

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Abstract

A human immunodeficiency virus type (HIV-1)-based lentiviral vector pseudotyped with the vesicular stomatitis virus envelope glycoprotein and encoding the GFP reporter gene was used to evaluate different methods of lentiviral vector titration. GFP expression, viral DNA quantification and the efficiency of vector DNA integration were assayed after infection of conventional HIV-1-permissive cell lines and human primary adult fibroblasts with the vector. We found that vector titers based on GFP expression determined by flow cytometry may vary by more than 50-fold depending on the cell type and the promoter-cell combination used. Interestingly, we observed that the viral integration process in primary HDFa cells was significantly more efficient compared to that in SupT1 or 293T cells. We propose that determination of the amount of integrated viral DNA by quantitative PCR be used in combination with the reporter gene expression assay.

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Acknowledgments

We thank Dr. D. Trono (University of Geneva, Switzerland) for providing us the pWPXLd and pCMV-dR8.91 plasmids, Dr. D. Nègre (ENS Lyon, France) for the pCMV-VSV-G plasmid and help for the production of vector preparation. We are grateful to the “Genetic analysis and vectorology” and “Flow cytometry” platforms of UMS3444 BioSciences Gerland Lyon Sud. VG was supported by a doctorate fellowship from Cluster 10 (Infectiology) of the Région Rhône-Alpes. We also thank Pr. P. Boulanger for critical reading of the manuscript.

Conflict of interest

The authors declare they have no conflict of interest.

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Authors and Affiliations

  1. Université de Lyon, 69000, Lyon, France

    Virginie Gay, Karen Moreau, Saw-See Hong & Corinne Ronfort

  2. Université Lyon 1, 69007, Lyon, France

    Virginie Gay, Karen Moreau, Saw-See Hong & Corinne Ronfort

  3. French National Institute for Agricultural Research (INRA), UMR754, Rétrovirus et Pathologie Comparée, 69007, Lyon, France

    Virginie Gay, Karen Moreau, Saw-See Hong & Corinne Ronfort

  4. UMS3444 BioSciences Gerland Lyon Sud, 69007, Lyon, France

    Virginie Gay, Karen Moreau, Saw-See Hong & Corinne Ronfort

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  1. Virginie Gay
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  2. Karen Moreau
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  3. Saw-See Hong
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  4. Corinne Ronfort
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Correspondence to Corinne Ronfort.

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Gay, V., Moreau, K., Hong, SS. et al. Quantification of HIV-based lentiviral vectors: influence of several cell type parameters on vector infectivity. Arch Virol 157, 217–223 (2012). https://doi.org/10.1007/s00705-011-1150-5

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  • DOI: https://doi.org/10.1007/s00705-011-1150-5

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